Dual luciferase reported gene assay kit

Reporter gene transfection and luciferase activity assay were performed as follows: cells in confluent growth on a 24 well plate were co-transfected with the firefly luciferase reporter (0.2 μg) along with the Renilla luciferase reporter (0.02 μg), which was used for normalization, using Attractene reagent (Qiagen, Hilden, Germany) according to the protocols provided by the manufacturers. The reporter plasmids of AP-1 were purchased from Biotime Biotechnology, China. The pAP1-Luc contains SV40 early enhancer/promoter. The AP-1 response element was listed as follows: GGCCTAACTG GCCGGTACCG CTAGCTGACT AATGACTAAT GACTAATGAC; CCGGATTGAC CGGCCATGGC GATCGACTGA TTACTGATTA CTGATTACTG. The luciferase activity was measured in cellular extracts using a dual luciferase reported gene assay kit (Promega, San Luis Obispo, CA, USA). The relative activity of the reporter gene was calculated by dividing the signals from firefly luciferase reporter by the signals obtained from Renilla luciferase reporter.

Reporter gene transfection and luciferase activity assay were performed as follow: cells on a 24 well plate were co-transfected with the firefly luciferase reporter (50 ng) along with the Renilla luciferase reporter (Promega) (20 ng) for 24 h using Lipofectamine 2000 (Thermo Fisher Scientific, America) according to the protocols provided by manufacturers. The reporter plasmid of TEAD was purchased from Addgene (Addgene, America). The luciferase activity was measured in cellular extracts using a dual luciferase reported gene assay kit (Promega). The relative activity of the reporter gene was calculated by dividing the signals from firefly luciferase reporter by the signals obtained from Renilla luciferase reporter.